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human intestinal organoid

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https://www.readbyqxmd.com/read/30526881/human-intestinal-organoids-maintain-self-renewal-capacity-and-cellular-diversity-in-niche-inspired-culture-condition
#1
Masayuki Fujii, Mami Matano, Kohta Toshimitsu, Ai Takano, Yohei Mikami, Shingo Nishikori, Shinya Sugimoto, Toshiro Sato
Cellular diversity that shapes tissue architecture and function is governed by multiple niche signals. Nonetheless, maintaining cellular diversity in human intestinal organoids has been challenging. Based on niche ligands present in the natural stem cell milieu, we establish a refined organoid culture condition for intestinal epithelia that allows human intestinal organoids to concurrently undergo multi-differentiation and self-renewal. High-throughput screening reveals that the combination of insulin-like growth factor 1 (IGF-1) and fibroblast growth factor 2 (FGF-2) enhances the clonogenic capacity and CRISPR-genome engineering efficiency of human intestinal stem cells...
December 6, 2018: Cell Stem Cell
https://www.readbyqxmd.com/read/30521806/small-heterodimer-partner-and-fibroblast-growth-factor-19-inhibit-expression-of-npc1l1-in-mouse-intestine-and-cholesterol-absorption
#2
Young-Chae Kim, Sangwon Byun, Sunmi Seok, Grace Guo, H Eric Xu, Byron Kemper, Jongsook Kim Kemper
BACKGROUND & AIMS: The nuclear receptor subfamily 0 group B member 2 (NR0B2, also called SHP) is expressed at high levels in liver and intestine. Postprandial fibroblast growth factor 19 (human FGF19, mouse FGF15) signaling increases the transcriptional activity of SHP. We studied the functions of SHP and FGF19 in intestines of mice, including their regulation of expression of the cholesterol transporter NPC1-like intracellular cholesterol transporter 1 (NPC1L1) and cholesterol absorption...
December 3, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30504008/high-throughput-single-cell-transcriptomics-on-organoids
#3
REVIEW
Agnieska Brazovskaja, Barbara Treutlein, J Gray Camp
Three-dimensional (3D) tissues grown in culture from human stem cells offer the incredible opportunity to analyze and manipulate human development, and to generate patient-specific models of disease. Methods to sequence DNA and RNA in single cells are being used to analyze these so-called 'organoid' systems in high-resolution. Single-cell transcriptomics has been used to quantitate the similarity of organoid cells to primary tissue counterparts in the brain, intestine, liver, and kidney, as well as identify cell-specific responses to environmental variables and disease conditions...
November 29, 2018: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/30488522/the-effect-of-ptc-mutations-on-cftr-mrna-abundance-in-human-nasal-epithelium-and-intestinal-organoids-a-basis-for-read-through-therapies-in-cystic-fibrosis
#4
Luka A Clarke, Nikhil T Awatade, Veronica M Felicio, Iris A Silva, Maite Calucho, Luisa Pereira, Pilar Azevedo, José Cavaco, Celeste Barreto, Carmen Bertuzzo, Silvia Gartner, Jeffrey Beekman, Margarida D Amaral
A major challenge in Cystic Fibrosis research is applying mutation-specific therapy to individual patients with diverse and rare CFTR genotypes. Read-through agents are currently the most promising approach for Class I mutations that introduce premature termination codons (PTCs) into CFTR mRNA. However, variations in degradation of PTC containing transcripts by nonsense mediated decay (NMD) might lower read-through efficacy. Allele specific quantitative real time (qRT-)PCR was used to measure variations in CFTR mRNA abundance for several PTC mutations in respiratory cells and intestinal organoids...
November 28, 2018: Human Mutation
https://www.readbyqxmd.com/read/30478383/dclk1-in-tuft-cells-promotes-inflammation-driven-epithelial-restitution-and-mitigates-chronic-colitis
#5
Jun Yi, Kirk Bergstrom, Jianxin Fu, Xindi Shan, J Michael McDaniel, Samuel McGee, Dongfeng Qu, Courtney W Houchen, Xiaowei Liu, Lijun Xia
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by defective intestinal barrier integrity toward the microbiota and epithelial damage. Double cortin-like kinase 1 (Dclk1), a marker of intestinal tuft cells, can regulate tissue regenerative responses, but its role in epithelial repair during bacterial-dependent chronic colitis is unclear. We addressed this question using our recently developed mouse model of spontaneous microbiota-dependent colitis induced by mucin-type O-glycan deficiency (DKO), which recapitulates most features of human UC...
November 26, 2018: Cell Death and Differentiation
https://www.readbyqxmd.com/read/30478287/an-upstream-protein-coding-region-in-enteroviruses-modulates-virus-infection-in-gut-epithelial-cells
#6
Valeria Lulla, Adam M Dinan, Myra Hosmillo, Yasmin Chaudhry, Lee Sherry, Nerea Irigoyen, Komal M Nayak, Nicola J Stonehouse, Matthias Zilbauer, Ian Goodfellow, Andrew E Firth
Enteroviruses comprise a large group of mammalian pathogens that includes poliovirus. Pathology in humans ranges from sub-clinical to acute flaccid paralysis, myocarditis and meningitis. Until now, all of the enteroviral proteins were thought to derive from the proteolytic processing of a polyprotein encoded in a single open reading frame. Here we report that many enterovirus genomes also harbour an upstream open reading frame (uORF) that is subject to strong purifying selection. Using echovirus 7 and poliovirus 1, we confirmed the expression of uORF protein in infected cells...
November 26, 2018: Nature Microbiology
https://www.readbyqxmd.com/read/30465020/nhe8-deficiency-promotes-colitis-associated-cancer-in-mice-via-expansion-of-lgr5-expressing-cells
#7
Hua Xu, Jing Li, Hao Chen, Fayez K Ghishan
Background & Aims: Lgr5 overexpression has been detected in colorectal cancers (CRCs), including some cases of colitis-associated CRCs. In colitis-associated CRCs, chronic inflammation is a contributing factor in carcinogenesis. We recently reported that intestinal Na+ /H+ exchanger isoform 8 (NHE8) plays an important role in intestinal mucosal protection and that loss of NHE8 expression results in an ulcerative colitis-like condition. Therefore, we hypothesized that NHE8 may be involved in the development of intestinal tumors...
2019: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/30443904/organ-on-chip-models-implications-in-drug-discovery-and-clinical-applications
#8
REVIEW
Rahul Mittal, Frank W Woo, Carlo S Castro, Madeline A Cohen, Joana Karanxha, Jeenu Mittal, Tanya Chhibber, Vasanti M Jhaveri
Before a lead compound goes through a clinical trial, preclinical studies utilize two-dimensional (2D) in vitro models and animal models to study the pharmacodynamics and pharmacokinetics of that lead compound. However, these current preclinical studies may not accurately represent the efficacy and safety of a lead compound in humans, as there has been a high failure rate of drugs that enter clinical trials. All of these failures and the associated costs demonstrate a need for more representative models of human organ systems for screening in the preclinical phase of drug development...
November 15, 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/30418409/bacteriophage-therapy-testing-against-shigella-flexneri-in-a-novel-human-intestinal-organoid-derived-infection-model
#9
Alejandro Llanos-Chea, Robert J Citorik, Kourtney P Nickerson, Laura Ingano, Gloria Serena, Stefania Senger, Timothy K Lu, Alessio Fasano, Christina S Faherty
OBJECTIVE: Enteric bacterial pathogens cause diarrheal disease and mortality at significant rates throughout the world, particularly in children under the age of 5 years. Our ability to combat bacterial pathogens has been hindered antibiotic resistance, a lack of effective vaccines, and accurate models of infection. With the renewed interest in bacteriophage therapy, we sought to use a novel human intestinal model to investigate the efficacy of a newly isolated bacteriophage against Shigella flexneri...
November 8, 2018: Journal of Pediatric Gastroenterology and Nutrition
https://www.readbyqxmd.com/read/30407734/human-milk-oligosaccharides-increase-mucin-expression-in-experimental-necrotizing-enterocolitis
#10
Richard You Wu, Bo Li, Yuhki Koike, Pekka Määttänen, Hiromu Miyake, Marissa Cadete, Kathene C Johnson-Henry, Steven R Botts, Carol Lee, Thomas R Abrahamsson, Eva Landberg, Agostino Pierro, Philip M Sherman
SCOPE: Necrotizing enterocolitis (NEC) is a leading cause of morbidity and death in preterm infants, occurring more often in formula-fed than breastfed infants. Studies in both rats and humans show that human milk oligosaccharides (HMOs) lower the incidence of NEC, but the mechanism underlying such protection is currently unclear. METHODS AND RESULTS: Here by extracting HMOs from pooled human breastmilk, we investigated the impact of HMOs on the intestinal mucin levels in a murine model of NEC...
November 8, 2018: Molecular Nutrition & Food Research
https://www.readbyqxmd.com/read/30365932/trib3-interacts-with-beta-catenin-and-tcf4-to-increase-stem-cell-features-of-colorectal-cancer-stem-cells-and-tumorigenesis
#11
Fang Hua, Shuang Shang, Yu-Wei Yang, Hai-Zeng Zhang, Tian-Lei Xu, Jiao-Jiao Yu, Dan-Dan Zhou, Bing Cui, Ke Li, Xiao-Xi Lv, Xiao-Wei Zhang, Shan-Shan Liu, Jin-Mei Yu, Feng Wang, Cheng Zhang, Bo Huang, Zhuo-Wei Hu
BACKGROUND & AIMS: Activation of Wnt signaling to beta-catenin contributes to development of colorectal cancer (CRC). Expression of tribbles pseudokinase 3 (TRIB3) is increased in some colorectal tumors and associated with poor outcome. We investigated whether increased TRIB3 expression promotes stem cell features of CRC cells and tumor progression by interacting with the Wnt signaling pathway. METHODS: We performed studies with C57BL/6J-ApcMin /J mice injected with an adeno-associated virus vector that expresses a small hairpin RNA against Trib3 mRNA (ApcMin /J-Trib3KD ) or a control vector (ApcMin /J-Ctrl)...
October 23, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30364784/dynamic-formation-of-microvillus-inclusions-during-enterocyte-differentiation-in-munc18-2-deficient-intestinal-organoids
#12
Mohammed H Mosa, Ophélie Nicolle, Sophia Maschalidi, Fernando E Sepulveda, Aurelien Bidaud-Meynard, Constantin Menche, Birgitta E Michels, Grégoire Michaux, Geneviève de Saint Basile, Henner F Farin
Background & Aims: Microvillus inclusion disease (MVID) is a congenital intestinal malabsorption disorder caused by defective apical vesicular transport. Existing cellular models do not fully recapitulate this heterogeneous pathology. The aim of this study was to characterize 3-dimensional intestinal organoids that continuously generate polarized absorptive cells as an accessible and relevant model to investigate MVID. Methods: Intestinal organoids from Munc18-2 / Stxbp2 -null mice that are deficient for apical vesicular transport were subjected to enterocyte-specific differentiation protocols...
2018: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/30344100/a-comprehensive-human-gastric-cancer-organoid-biobank-captures-tumor-subtype-heterogeneity-and-enables-therapeutic-screening
#13
Helen H N Yan, Hoi Cheong Siu, Simon Law, Siu Lun Ho, Sarah S K Yue, Wai Yin Tsui, Dessy Chan, April S Chan, Stephanie Ma, Ka On Lam, Sina Bartfeld, Alice H Y Man, Bernard C H Lee, Annie S Y Chan, Jason W H Wong, Priscilla S W Cheng, Anthony K W Chan, Jiangwen Zhang, Jue Shi, Xiaodan Fan, Dora L W Kwong, Tak W Mak, Siu Tsan Yuen, Hans Clevers, Suet Yi Leung
Gastric cancer displays marked molecular heterogeneity with aggressive behavior and treatment resistance. Therefore, good in vitro models that encompass unique subtypes are urgently needed for precision medicine development. Here, we have established a primary gastric cancer organoid (GCO) biobank that comprises normal, dysplastic, cancer, and lymph node metastases (n = 63) from 34 patients, including detailed whole-exome and transcriptome analysis. The cohort encompasses most known molecular subtypes (including EBV, MSI, intestinal/CIN, and diffuse/GS, with CLDN18-ARHGAP6 or CTNND1-ARHGAP26 fusions or RHOA mutations), capturing regional heterogeneity and subclonal architecture, while their morphology, transcriptome, and genomic profiles remain closely similar to in vivo tumors, even after long-term culture...
October 13, 2018: Cell Stem Cell
https://www.readbyqxmd.com/read/30305617/lrh-1-mitigates-intestinal-inflammatory-disease-by-maintaining-epithelial-homeostasis-and-cell-survival
#14
James R Bayrer, Hongtao Wang, Roy Nattiv, Miyuki Suzawa, Hazel S Escusa, Robert J Fletterick, Ophir D Klein, David D Moore, Holly A Ingraham
Epithelial dysfunction and crypt destruction are defining features of inflammatory bowel disease (IBD). However, current IBD therapies targeting epithelial dysfunction are lacking. The nuclear receptor LRH-1 (NR5A2) is expressed in intestinal epithelium and thought to contribute to epithelial renewal. Here we show that LRH-1 maintains intestinal epithelial health and protects against inflammatory damage. Knocking out LRH-1 in murine intestinal organoids reduces Notch signaling, increases crypt cell death, distorts the cellular composition of the epithelium, and weakens the epithelial barrier...
October 10, 2018: Nature Communications
https://www.readbyqxmd.com/read/30273559/epithelial-rnase-h2-maintains-genome-integrity-and-prevents-intestinal-tumorigenesis-in-mice
#15
Konrad Aden, Kareen Bartsch, Joseph Dahl, Martin A M Reijns, Daniela Esser, Raheleh Sheibani-Tezerji, Anupam Sinha, Felix Wottawa, Go Ito, Neha Mishra, Katharina Knittler, Adam Burkholder, Lina Welz, Johan van Es, Florian Tran, Simone Lipinski, Nassim Kakavand, Christine Boeger, Ralph Lucius, Witigo von Schoenfels, Clemens Schafmayer, Lennart Lenk, Athena Chalaris, Hans Clevers, Christoph Röcken, Christoph Kaleta, Stefan Rose-John, Stefan Schreiber, Thomas Kunkel, Björn Rabe, Philip Rosenstiel
BACKGROUND & AIMS: RNase H2 is a holoenzyme comprising 3 subunits (ribonuclease H2 subunits A, B, and C) that cleaves RNA:DNA hybrids and removes misincorporated ribonucleotides from genomic DNA via ribonucleotide excision repair. Ribonucleotide incorporation by eukaryotic DNA polymerases occurs during every round of genome duplication and produces the most frequent type of naturally occurring DNA lesion. We investigated whether intestinal epithelial proliferation requires RNase H2 function and whether RNase H2 activity is disrupted during intestinal carcinogenesis...
September 28, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30273442/dual-specificity-phosphatase-6-deletion-protects-the-colonic-epithelium-against-inflammation-and-promotes-both-proliferation-and-tumorigenesis
#16
Katia Beaudry, Marie-Josée Langlois, Amélie Montagne, Sébastien Cagnol, Julie C Carrier, Nathalie Rivard
The Ras/mitogen-activated protein kinase (MAPK) pathway controls fundamental cellular processes such as proliferation, differentiation, and apoptosis. The dual-specificity phosphatase 6 (DUSP6) regulates cytoplasmic MAPK signaling by dephosphorylating and inactivating extracellular signal-regulated kinase (ERK1/2) MAPK. To determine the role of DUSP6 in the maintenance of intestinal homeostasis, we characterized the intestinal epithelial phenotype of Dusp6 knockout (KO) mice under normal, oncogenic, and proinflammatory conditions...
October 1, 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/30267714/intestinal-inflammation-and-dysregulated-immunity-in-patients-with-inherited-caspase-8-deficiency
#17
Anna S Lehle, Henner F Farin, Benjamin Marquardt, Birgitta E Michels, Thomas Magg, Yue Li, Yanshan Liu, Maryam Ghalandary, Katja Lammens, Sebastian Hollizeck, Meino Rohlfs, Fabian Hauck, Raffaele Conca, Christoph Walz, Batia Weiss, Atar Lev, Amos J Simon, Olaf Groß, Moritz M Gaidt, Veit Hornung, Hans Clevers, Nadine Yazbeck, Rima Hanna-Wakim, Dror S Shouval, Neil Warner, Raz Somech, Aleixo M Muise, Scott S Snapper, Philip Bufler, Sibylle Koletzko, Christoph Klein, Daniel Kotlarz
Caspase-8 (CASP8) is a protease that initiates apoptosis and regulates inflammation and immune responses. We identified germline mutations in CASP8 in 3 unrelated patients with infant-onset inflammatory bowel disease: 2 patients were homozygous for the mutation 710A>G, p.Q237R, which resulted in reduced protein expression, and 1 patient carried the mutation 793C>T, p.R265W. We isolated peripheral blood mononuclear cells from our index patient and observed defects in T- and B-cell maturation, proliferation, and/or activation...
September 26, 2018: Gastroenterology
https://www.readbyqxmd.com/read/30265443/initiation-expansion-and-cryopreservation-of-human-primary-tissue-derived-normal-and-diseased-organoids-in-embedded-three-dimensional-culture
#18
James Clinton, Penney McWilliams-Koeppen
Organoids are primary patient-derived micro tissues grown within a three-dimensional extracellular matrix that better represents in vivo physiology and genetic diversity than existing two-dimensional cell lines. Organoids rely on the self-renewal and differentiation of tissue-resident stem cells that expand in culture and self-organize into complex three-dimensional structures. Depending on the tissue, organoids typically lack stromal, vascular, neural, and immune cells but otherwise can contain cells from all the respective tissue-specific cell lineages found in vivo...
September 28, 2018: Current Protocols in Cell Biology
https://www.readbyqxmd.com/read/30261339/cellulose-based-scaffolds-for-fluorescence-lifetime-imaging-assisted-tissue-engineering
#19
Neil O'Donnell, Irina A Okkelman, Peter Timashev, Tatyana I Gromovykh, Dmitri B Papkovsky, Ruslan I Dmitriev
Quantitative measurement of pH and metabolite gradients by microscopy is one of the challenges in the production of scaffold-grown organoids and multicellular aggregates. Herein, we used the cellulose-binding domain (CBD) of the Cellulomonas fimi CenA protein for designing biosensor scaffolds that allow measurement of pH and Ca2+ gradients by fluorescence intensity and lifetime imaging (FLIM) detection modes. By fusing CBD with pH-sensitive enhanced cyan fluorescent protein (CBD-ECFP), we achieved efficient labeling of cellulose-based scaffolds based on nanofibrillar, bacterial cellulose, and decellularized plant materials...
September 25, 2018: Acta Biomaterialia
https://www.readbyqxmd.com/read/30259138/developing-a-3d-intestinal-epithelium-model-for-livestock-species
#20
Hayley Derricott, Lisa Luu, Wai Yee Fong, Catherine S Hartley, Luke J Johnston, Stuart D Armstrong, Nadine Randle, Carrie A Duckworth, Barry J Campbell, Jonathan M Wastling, Janine L Coombes
The in vitro 3D culture of intestinal epithelium is a valuable resource in the study of its function. Organoid culture exploits stem cells' ability to regenerate and produce differentiated epithelium. Intestinal organoid models from rodent or human tissue are widely available whereas large animal models are not. Livestock enteric and zoonotic diseases elicit significant morbidity and mortality in animal and human populations. Therefore, livestock species-specific models may offer novel insights into host-pathogen interactions and disease responses...
September 26, 2018: Cell and Tissue Research
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